GSM7011356: Budding yeast, Rec8 ChIP, 8h in meiosis, rep1; Saccharomyces cerevisiae; ChIP-Seq
From a synchronous meiotic culture, 50 ml of samples were collected at various time points and crosslinked using formaldehyde (1%) for 30 min. Cells were lysed using Beadbeater (Unigenetics). The cell extract was sonicated using Bioruptor (Diagenode) and the debris was removed by centrifugation. Protein A agarose beads were used to preclear the cell lysate from each sample. To the pre-cleared lysate, 2.5 μl Rec8 anti-serum and 7.5 mg of Protein-A agarose beads blocked with BSA were added. After incubation, the lysate was removed by centrifugation, and beads were washed to remove non-specific binding. The immunoprecipitated material was eluted from the beads and decrosslinked along with RNase A treatment. The DNA was purified following Proteinase K treatment using QIAGEN PCR purification kit. ChIP-seq libraries were prepared using the illumina ChIP-Seq TruSeq kit from 20 ng of DNA as starting material for input and IP samples.